Raman spectroscopy is a technique for performing chemical analysis. High intensity monochromatic light, such as that provided by a laser, is directed onto an analyte molecule (or sample) that is to be chemically analyzed. A majority of the incident photons are elastically scattered by the analyte molecule, wherein the scattered photons have the same energy (and, therefore, the same frequency) as the incident photons. This elastic scattering is termed Rayleigh scattering, and the elastically scattered photons and radiation are termed Rayleigh photons and Rayleigh radiation, respectively. However, a small fraction of the photons (e.g., about 1 in 107 photons) are inelastically scattered by the analyte molecules. These inelastically scattered photons have a different frequency than the incident photons. This inelastic scattering of photons is termed the Raman effect. The inelastically scattered photons may have frequencies greater than, or, more typically, less than the frequency of the incident photons.
When an incident photon collides with a molecule, energy may be transferred from the photon to the molecule or from the molecule to the photon. When energy is transferred from the photon to the molecule, the scattered photon will emerge from the sample having a lower energy and a corresponding lower frequency. These lower-energy Raman scattered photons are commonly referred to in Raman spectroscopy as the Stokes radiation. A small fraction of the analyte molecules are already in an energetically excited state. When an incident photon collides with an excited molecule, energy may be transferred from the molecule to the photon, which will emerge from the sample having a higher energy and a corresponding higher frequency. These higher-energy Raman scattered photons are commonly referred to in Raman spectroscopy as the anti-Stokes radiation.
The Stokes and the anti-Stokes radiation is detected by a detector, such as a photomultiplier or a wavelength-dispersive spectrometer, which converts the energy of the impinging photons into an electrical signal. The characteristics of the electrical signal are at least partially a function of the energy (or wavelength, frequency, wave number, etc.) of the impinging photons and the number of the impinging photons per unit time (intensity). The electrical signal generated by the detector can be used to produce a spectral graph of intensity as a function of frequency for the detected Raman signal (i.e., the Stokes and anti-Stokes radiation). A unique Raman spectrum corresponding to the particular analyte may be obtained by plotting the intensity of the inelastically scattered Raman photons against their frequency or, equivalently and more commonly, their wavenumber in units of inverse centimeters. This unique Raman spectrum may be used for many purposes such as identifying an analyte, identifying chemical states or bonding of atoms and molecules in the analyte, and determining physical and chemical properties of the analyte. Raman spectroscopy may be used to analyze a single molecular species or mixtures of different molecular species. Furthermore, Raman spectroscopy may be performed on a number of different types of molecular configurations, such as organic and inorganic molecules in either crystalline or amorphous states.
Molecular Raman scattering of photons is a weak process. As a result, powerful, costly laser sources typically are used to generate high intensity excitation radiation to increase the weak Raman signal for detection. Surface enhanced Raman spectroscopy (SERS) is a technique that allows for generation of a stronger Raman signal from an analyte relative to non-SERS Raman spectroscopy for a sample with the same number of analyte molecules. In SERS, the analyte molecules are adsorbed onto, or placed adjacent to, an activated metal surface or structure, termed herein a SERS-active structure. The interactions between the molecules and the surface cause an increase in the strength of the Raman signal. Several SERS-active structures have been employed in SERS techniques, including activated electrodes in electrolytic cells, activated metal colloid solutions, and activated metal substrates such as a roughened metal surface or metal islands formed on a substrate. For example, it has been shown that adsorbing analyte molecules onto or near a specially roughened metal surface made from silver or gold may enhance the Raman scattering intensity by factors of between 103 and 106.
As used herein, SERS-active material refers to the metallic material (or other material that can be found to have a similar effect) that is distributed across the substrate to form the SERS-active structures. As used herein in the context of SERS-active material, nanosection refers to any particular localized section of SERS-active material, such as an island of SERS-active material, a colloidal particle of SERS-active material, or other localized segment such as may be associated with a particular surface bump or portion of a differently shaped surface. Thus, any particular SERS-active structure comprises a patterned arrangement (which can be random, pseudorandom, or ordered) of nanosections of SERS-active material (which can be uniformly shaped or highly non-uniform and irregular in shape).
Although theories such as electromagnetic enhancement and chemical enhancement are set forth in the literature, the mechanism of Raman signal enhancement exhibited in SERS is not completely understood. Empirically, it has been observed that the intensification of the Raman scattered photons for any particular wavelength is only provided for analyte molecules in the vicinity of certain localized structures, bumps, or cavities formed by the SERS-active material (i.e., in the vicinity of certain particular patterns/shapes of SERS-active material nanosections) with sizes varying between 1 nm and several hundreds of nanometers, known as hot spots. The existence of a particular hot spot can be volatile to even small changes in any of a variety of conditions, including the local patterns/shapes of SERS-active nanosections, the position of the analyte molecule(s) relative to that location, the type of analyte molecule(s), the type of SERS-active material, and the wavelength of the incident light. In view of this volatility, it would be desirable to provide a SERS system in which increased Raman intensification can be achieved for a greater variety of analyte molecules and for greater variations in incident wavelengths, system materials, and other field conditions associated with the molecular sensing process. Other issues arise as would be apparent to one skilled in the art upon reading the present disclosure.